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Characterization of a novel mechanism restricting virus infection in reproductive tissues

Objective

Like mammals, insects are infected by many viruses. Among them, arthropod-borne viruses are an increasing worldwide health concern. Insects have potent innate antiviral defenses, of which RNA interference (RNAi) is the main and best studied. In the fruitfly Drosophila melanogaster, the siRNA pathway controls viral replication in somatic tissues. The piRNA pathway, another RNAi based response, acts specifically in the reproductive tract (germline and follicular cells (FC), a monolayer of somatic cells surrounding the germline), to protect the genome against transposon mobilization. Other innate immunity or stress pathways also contribute to the antiviral defense.
The host laboratory obtained evidence for a new mechanism controlling viral replication in the FCs of Drosophila. Indeed, a viral replicon derived from Flock House Virus and expressing GFP (FHVΔB2-GFP) is completely derepressed in somatic tissues of mutants for the siRNA pathway, except in FCs, where derepression is partial. No piRNAs of viral origin can be detected in these cells. This viral replicon provides a unique system to decipher a novel pathway restricting viral replication.
For this, I will use a combination of forward and reverse genetic screens. EMS mutagenesis, screening for GFP expression and genome resequencing will be used to uncover genes responsible for restricting the replicon in FCs. Next, I will use GFP expression to sort restrictive and permissive FCs in a siRNA pathway mutant followed by RNA sequencing to identify differentially expressed genes. The function of the identified genes will be tested in vivo, using shRNA Drosophila lines. Finally, OSS cells, an FC-derived cell line, will be transformed with the FHVΔB2-GFP replicon to evaluate candidate gene function ex vivo.
The identification and characterization of genes involved in a novel viral restriction pathway will increase the knowledge about innate antiviral immunity, an important scientific topic with human health implications.

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MSCA-IF-EF-ST - Standard EF

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Call for proposal

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(opens in new window) H2020-MSCA-IF-2015

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Coordinator

CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE CNRS
Net EU contribution

Net EU financial contribution. The sum of money that the participant receives, deducted by the EU contribution to its linked third party. It considers the distribution of the EU financial contribution between direct beneficiaries of the project and other types of participants, like third-party participants.

€ 185 076,00
Total cost

The total costs incurred by this organisation to participate in the project, including direct and indirect costs. This amount is a subset of the overall project budget.

€ 185 076,00
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